This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We will use the proposed LAMMP Spatially Modulated Microbeams (SMM) system to measure mechanical changes in the extra cellular matrix (ECM) in response to MT1-MMP expressing cells. In our first aim, adhesion forces between extracellular matrix (ECM) proteins and membrane receptors on the surface of MT1-MMP expressing cells will be measured. Microbeads coated with specific ECM proteins we be held in an optical force clamp to measure adhesion forces. A FRET based sensor of MT1-MMP activity on the cell surface will monitor MT1-MMP activity levels, which will be correlated in time to adhesion strength. In our second aim, MT1-MMP expressing cells will be cultured within ECM protein hydrogels containing various mixtures of ECM protein types. By pulling and measuring displacements of multiple microbeads embedded within the hydrogel, we will track changes in mechanical properties of the gel proximal and distal to cells with varying expression levels in response to chemical stimulation. This collaboration will offer new insights into the kinetics of ECM digestion by membrane-bound MT1-MMP.